Experimental Design

Hyperlipidemia Group: Giving normal drinking water. Fed with 50g of 1.5% cholesterol enriched diet in the morning and 90-120g of ordinary diet in the evening.

OLE Group: Giving normal drinking water. Fed with 50g of 1.5% cholesterol enriched diet together with 50ml of OLE every morning; and 90-120g of ordinary diet in the evening.

Measuring Parameters

The level of total cholesterol (CH), Triglyceride (TG), High Density Lipoprotein (HDL), and Superoxide Dismutase (SOD) from Rabbit Auricular Veins was measured on the 2nd week, 4th week and 7th week of the experiment to monitor the changes of lipid profile and self protecting mechanism in the experimental animals.

At the end of the 23rd week, the thoracic aorta of the rabbit was taken out and preceded with fixing, staining as well as analyzing with an image analyzer. This was to measure the area and percentage of intravascular atherosclerosis plaque to observe the development of intravascular atherosclerosis plaque.

1.2 Result (ANOVA Analysis)

1.2.1 CH (mg/dl)

The elevation of serum total cholesterol level in OLE group reduced 970% compared to Hyperlipidemia group. Hence, this has shown that OLE can help to restrain the elevation of serum cholesterol level. After drinking OLE for one week, serum cholesterol level in OLE group had reduced apparently compared to Hyperlipidemia group and became more apparent after the second week. Thus, this has shown that OLE can help to lower serum cholesterol level at the early state of Hyperlipidemia. (Refer Table 1)

Table 1: Comparison of Serum Cholesterol Level among the Three Groups

X±SE

Control Group

Hyperlipidemia Group

OLE Group

1st Week

90.13±14.32

127.86±46.19

66.64±9.9

2nd Week

51.9±11.7

135.7±29.9

83.5±15.9

1.2.2 TG (mg/dl)

The elevation of serum triglyceride level in OLE group reduced 120% compared to Hyperlipidemia group. Hence, this has shown that OLE can help to restrain the elevation of triglyceride. After drinking OLE for one week, triglyceride level in OLE group had reduced apparently compared to Hyperlipidemia group, and became more apparent after the third week. (Refer Table 2)

Table 2: Comparison of Serum Triglyceride Level among the Three Groups

X±SE

Control Group

Hyperlipidemia Group

OLE Drink Group

2nd Week

63.1±12.5

69.8±11.4

62.1±5.9

4th Week

64.7±11.9

113.0±39.1

70.2±14.3

7th Week

60.4±8.8

122.3±47.2

72.3±20.7

1.2.3 HDL (mg/dl)

Serum HDL level in OLE group had elevated compared to Hyperlipidemia group and became apparent on the 4th week of experiment. (Refer Table 3)

Table 3: Comparison of HDL Level among the Three Groups

X±SE

Control Group

Hyperlipidemia Group

OLE Drink Group

2nd Week

27.3±2.6

35.1±2.8

37.8±2.6

4th Week

31.4±4.4

34.6±3.7

64.9±29.7

1.2.4 SOD (NU/ML)

On the first week of the experiment, SOD level in OLE group was the highest among the three groups. (Refer Table 4)

Table 4: Comparison of SOD Level among the Three Groups

X±SE

Control Group

Hyperlipidemia Group

OLE Group

2nd Week

447.3±23.9

448.0±13.6

493.0±52.8

4th Week

497.4±27.9

446.3±11.9

516.2±7.7

The average increase of SOD level in OLE group was 20% higher than that of Hyperlipidemia group.

1-2.5 The Comparison of the Percentage Increase of CH, TG and SOD in OLE group, Hyperlipidemia and Normal Group

OLE Group

Hyperlipidemia

CH

1220%

2190%

TG

90%

210%

SOD

30%

10%

Based on the results above, OLE effectively inhibits CH and TG in serum from rising, and at the same time, increases SOD levels, which prevents the occurrence and development of atherosclerosis to a certain extent. Therefore, long-term and regular consumption of OLE is an effective way in preventing atherosclerosis. Besides, the experiments results have shown that the body weights of the OLE group decreased as compared to the Controls and Hyperlipidemia group. This might due to the inhibition of body fat accumulation.

1-3 Vascular atherosclerosis plaque area and the percentage of it out of the total intimal area in each group. Observations shown in Figure 1, 2, 3.

Figure 1: was the formation of thoracic aortic intima atherosclerosis in OLE group, in which its area constituted 41% out of the total intimal area.

Figure 2: was the formation of thoracic aortic intima atherosclerosis in Hyperlipidemia group, in which its area constituted 97% out of the total intimal area.

Figure 3: was the formation of thoracic aortic intima atherosclerosis in the Controls, in which its area constituted 4% out of the total intimal area.

Results analyzed from the figures were identical with the results obtained from CH, TG and HDL levels.

The experiments indicated that continual feeding of rabbits with 1.5% of high cholesterol diet for 23 weeks, a significant result was shown: the intimal atherosclerosis plaque area of the Hyperlipidemia group exceeded 230% as compared with the OLE group. This has shown that OLE is miraculously effective in preventing atherosclerosis.

1.2 Methodology
1.2.1 For determination of resistance of cold stimulation ability, each mouse was fed with 0.5ml of OLE daily, and given normal water intake condition. Each of the mice in normal Controls was fed with 0.5ml of normal water daily, and was also given normal water intake condition.

2. Experiment and Experimental Results

2.1 Determination of resistance of cold stimulation ability: The mice were all fed with the different drinks as stated above. 1 hour later, all mice were put into 13oC water to swim; the survival time was then measured.
Each group of mice were let to swim in the different lanes in the different sizes of buckets. Survival time was measured. The water depth, surface diameter and water temperature were 20cm, 30cm and 13oC, respectively.
Results were recorded based on the average±SD of the survival time in the table below, using Minute as time unit.

Survival Time (m) /Group

Control

OLE Group

Single-ingredient 1 Group

8.2±8.0

16±1.0

14.2±0.90

P<0.01
Based on the results, it can be seen that both OLE and Single-ingredient 1 had significantly increased the ability in resisting cold stimulation. There is no significant difference in the other groups with other ingredients and Control.

2.2 Determination of static endurance in mice: 3 hours before the third day to be fed with different groups of drinks, the mice were being fed with the drinks mentioned above, and these mice were suspended on wires with 0.5mm in width and1 meter in length, 20cm from ground. Duration (seconds) of mice remained in suspended state was measured. This duration was the time where the mice grasped the wire from the beginning until they fell down. The duration in suspended state was recorded 3 times for each mouse. The sums of the average time of the different groups were recorded in the table below.

Duration in suspended state (s)/Group

Control

Single-ingredient 1 Group

OLE Group

51±4.0

63±5.0

70±6.0

Based on the results, OLE significantly increased the static endurance ability in mice. Its role was greater than that of Single-ingredient. This result ran parallel with the weightlifting and four hundred meters run results in China. It could be observed that taking a Single-ingredient can enhance athletes’ performances. This has shown that OLE in improving athletes’ performances during a competition is safe and feasible.

3. OLE is a non-toxic and non-side effect beverage and safe for long-term consumption.

According to the Procedures for Assessment of Food Safety, mice were tested in acute toxicity test, gene mutation test, mice bone marrow micronucleus test and mice sperm teratogenicity study. All the studies have shown that OLE is non-toxic and safe for consumption.